Self Quenching

This is perhaps not surprising since the dimer is shown to undergo efficient internal quenching. 

In order to further increase the solubility of the dye we are now synthesising tetra sulphonated derivates. We hope that the increased charge may besides., decrease the self quenching. If this is not the case then more Fe (II) will have to be added in order to trap Th this will lead to a reduction in the cell voltage through the Nernst term. 

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The experimental principle is illustrated in Fig. 3. The interaction of the polymer with the liposomal membranes causes the perturbation of the bilayer. This perturbation follows the leakage of calcein from the liposome. Calcein in high concentration in the liposome is self-quenched, but has strong fluorescence intensity by the leak from the liposome. Therefore, the extent of the membrane interaction can be estimated quantitatively from the fluorescence spectroscopy. 

In the equation, the subscripts 1 and 2 refer to the reference compound and the compound of interest, respectively, is the intensity of the fluorescent signal of each compound measured as peak height in centimeters, 8 is the molar absorptivity, c is the concentration in moles per liter, and is the fluorescence quantum yield. In this application, i is set at 1.00. The concentrations of the solutions that were tested ranged from 10 to 10 M. The solutions run at the higher concentrations were all checked for self-quenching, but none was found. All measurements, except the fluorescence-versus-solvent study, were made in 0.1-N phosphate buffer, pH 7.4. Slit settings on the Perkin-Elmer MPF-2A were 10 mp (nm) for both emission and excitation monochromators. 

Fig. 6.20. Types of assays to visualize lipid trafficking in membranes. (A) Self-quenching method. Here the self-quenching is released upon transfer to unlabeled acceptor membranes that are usually in large excess. (B) FRET assays. Here the donor membrane contains a transferable lipid (green) that is quenched by FRET to a non transferable acceptor lipid (red). Upon transfer to an unlabeled acceptor membrane the green-labeled lipid becomes unquenched. 

In Figure 5, the normalized emission spectra of the two solid hybrid materials, GFP/SBA-15 and GFP/Aerosil , are reported. The shape of the emission profile for GFP/SBA-15 follows closely that of the GFP in buffer solution, whereas the photoemission intensity of GFP/Aerosil is one order of magnitude lower and slightly different in its tale shape (spectra at the actual intensities not reported). This reduction in intensity could be explained by a multilayer arrangement of the protein molecules on the amorphous nanoparticles, which would explain both the difference in emission spectra ("self-quenching effect") and the difference in adsorption amount shown above. 

Once again, the most recent developments have been in the area of green phosphorescent materials where phenomenal efficiencies are now beginning to be coupled with good device lifetimes. The prototypical emitter of this type is iridium-tris-2-phenylpyridine (Ir(ppy)3) used as a dopant (Scheme 3.98), usually diluted into a carbazole type host, because it is prone to serious self-quenching problems. 

Poly(methyl methacrylate) [PMMA] is an excellent polymer for studying photoresist dissolution because of its minimal swelling characteristic. For this work, PMMA molecules were labelled with phenanthrene (Phe) dye since its fluorescence is quenched by MEK. In addition, this dye has the advantage of forming few excimers (23-241 which results in self-quenching. Thus, the reduction in fluorescence intensity of PMMA-Phe is virtually solely due to MEK quenching. Consequently, the permeation of MEK into a PMMA film can be monitored from fluorescence intensity decay. 

The value of the quantum yield of fluorescence of TIN in the PMMA film (calculated using the total film absorbance at the excitation wavelength) decreases from 1.2 x 10-3 to 5.0 x 10"4 when the concentration of TIN is increased from 0.07 mole% to 5.0 mole%. This suggests that the TIN molecules are involved in a concentration-dependent, self-quenching process. 

The dependence of the fluorescence quantum yields and lifetimes of these stabilizers on the nature of the solvent suggests that the excited-state, non-radiative processes are affected by solvation. In polar, hydroxylic solvents, values of the fluorescence quantum yield for the non proton-transferred form are significantly lower, and the fluorescence lifetimes are shorter, than those calculated for aprotic solvents. This supports the proposal of the formation, in alcoholic solvents, of an excited-state encounter complex which facilitates ESIPT. The observed concentration dependence of the fluorescence lifetime and intensity of the blue emission from TIN in polymer films provides evidence for a non-radiative, self-quenching process, possibly due to aggregation of the stabilizer molecules. 

Fig. 4 Illustration of homo-FRET and self-quenching by dimers, (a) The dyes (D) exchange their excitation energies and are bright emitters, (b) The presence of nonfluorescent dye dimer (DD) acting as the trap for excitation energy results in quenching of all the dyes located within FRET distance to it...

Since the same dye molecules can serve as both donors and acceptors and the transfer efficiency depends on the spectral overlap between the emission spectrum of the donor and the absorption spectrum of the acceptor, this efficiency also depends on the Stokes shift [53]. Involvement of these effects depends strongly on the properties of the dye. Fluoresceins and rhodamines exhibit high homo-FRET efficiency and self-quenching pyrene and perylene derivatives, high homo-FRET but little self-quenching and luminescent metal complexes may not exhibit homo-FRET at all because of their very strong Stokes shifts. 

The fluorescence quenching occurs when dye molecules are close to the metal. The energy from the first excited fluorophores can be consumed through a non-radiative path to the metal. A spacing layer is usually required to avoid this energy transfer process. In addition, the concentration of the dispersed dye molecules should be suitable to avoid self quenching [34, 81]. 

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